A highly Stable and Nonintegrated Human Artificial Chromosome (HAC) Containing the 2.4 Mb Entire Human Dystrophin Gene
A highly Stable and Nonintegrated Human Artificial Chromosome (HAC) Containing the 2.4 Mb Entire Human Dystrophin Gene
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DOI：
10.1038/mt.2008.253
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发表时间：
2009-02-01
影响因子：
12.4
通讯作者：
Oshimura, Mitsuo
中科院分区：
文献类型：
--
作者：
Hoshiya, Hidetoshi;Kazuki, Yasuhiro;Oshimura, Mitsuo
附加型载体具有递送包含所有关键调控元件的大基因的能力，是基因治疗的理想载体。人类人工染色体（HAC）具有将极大的遗传区域递送到宿主细胞而不整合到宿主基因组中的能力，从而防止可能的插入诱变和基因组不稳定性。Duchenne肌营养不良症（DMD）是由非常大的肌营养不良蛋白基因（2.4 Mb）突变引起的。我们在此报告了一种HAC载体的发展，该载体含有完整的人肌营养不良蛋白基因（DYS-HAC），该基因在小鼠和人永生化间充质干细胞（hiMSC）中稳定维持。将DYS-HAC转移到小鼠胚胎干（ES）细胞中，由ES细胞产生的嵌合小鼠中DYS-HAC衍生的人肌营养不良蛋白的同种型以组织特异性方式正确表达。因此，该HAC载体含有完整的肌营养不良蛋白基因及其天然调控元件，预计对DMD的未来基因和细胞治疗非常有用。
Episomal vector with the capacity to deliver a large gene containing all the critical regulatory elements is ideal for gene therapy. Human artificial chromosomes ( HACs) have the capacity to deliver an extremely large genetic region to host cells without integration into the host genome, thus preventing possible insertional mutagenesis and genomic instability. Duchenne muscular dystrophy (DMD) is caused by mutation in the extremely large dystrophin gene (2.4 Mb). We herein report the development of a HAC vector containing the entire human dystrophin gene (DYS-HAC) that is stably maintained in mice and human immortalized mesenchymal stem cells (hiMSCs). The DYS-HAC was transferred to mouse embryonic stem (ES) cells, and isoforms of the DYS-HAC-derived human dystrophin in the chimeric mice generated from the ES cells were correctly expressed in tissue-specific manner. Thus, this HAC vector containing the entire dystrophin gene with its native regulatory elements is expected to be extremely useful for future gene and cell therapies of DMD.